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. 2020 Jun 3;6(23):eaba0745. doi: 10.1126/sciadv.aba0745

Fig. 1. PVSRIPO translation persists in the presence of PKR-induced p-eIF2α(S51).

Fig. 1

(A) HeLa cells were transfected with control small interfering RNA (siRNA) or siRNAs targeting PKR or PERK (48 hours) and infected with PVSRIPO at a multiplicity of infection (MOI) of 10. At each interval shown, the cells were treated with puromycin (10 μM; 8 min) and harvested for immunoblot analysis of eIF2α status, host translation (puromycylated polypeptides), viral translation (2C, 2BC, P2), or cell death (PARP cleavage; *, cleavage fragment). (n = 3). (B) HeLa cells were infected with PVSRIPO and treated with vehicle or ISRIB (+), puromycylated as described for (A) and lysed at the indicated time points. (n = 3). (C) The biological effect of ISRIB in the assay shown in (B) was validated in HeLa cells treated with thapsigargin as shown.